Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Photoluminescence: Applications01:14

Photoluminescence: Applications

515
Photoluminescence offers a wide range of applications due to its inherent sensitivity and selectivity. This technique allows for both direct and indirect analyses of the analyte. Direct quantitative analysis is possible when the analyte exhibits a favorable quantum yield for fluorescence or phosphorescence. However, an indirect analysis may be feasible if the analyte is not fluorescent or phosphorescent, or if the quantum yield is unfavorable. Indirect methods include reacting the analyte with...
515
Labeling DNA Probes03:31

Labeling DNA Probes

8.4K
DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...
8.4K

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Myeloperoxidase and Eosinophil Peroxidase Inhibit Endotoxin Activity and Increase Mouse Survival in a Lipopolysaccharide Lethal Dose 90% Model.

Journal of immunology research·2019
Same author

Mechanism of Microbicidal Action of E-101 Solution, a Myeloperoxidase-Mediated Antimicrobial, and Its Oxidative Products.

Infection and immunity·2019
See all related articles

Related Experiment Video

Updated: Sep 29, 2025

Bioluminescence Imaging of NADPH Oxidase Activity in Different Animal Models
10:42

Bioluminescence Imaging of NADPH Oxidase Activity in Different Animal Models

Published on: October 22, 2012

16.4K

Haloperoxidase-Catalyzed Luminol Luminescence.

Robert C Allen1

  • 1Department of Pathology, Creighton University, Omaha, NE 68178, USA.

Antioxidants (Basel, Switzerland)
|March 25, 2022
PubMed
Summary

Luminol luminescence reveals differences in peroxidase enzyme kinetics. Horseradish peroxidase (HRP) kinetics differ from myeloperoxidase (MPO) and eosinophil peroxidase (EPO) kinetics, especially concerning hydrogen peroxide concentration and halide requirements.

Keywords:
chemiluminescenceeosinophil peroxidasehalide oxidationhaloperoxidasehorseradish peroxidasekinetic analysisluminol luminescencemyeloperoxidasereaction ordersinglet molecular oxygen

More Related Videos

Fast and Specific Assessment of the Halogenating Peroxidase Activity in Leukocyte-enriched Blood Samples
05:17

Fast and Specific Assessment of the Halogenating Peroxidase Activity in Leukocyte-enriched Blood Samples

Published on: July 28, 2016

10.4K
A Novel Technique for Generating and Observing Chemiluminescence in a Biological Setting
08:57

A Novel Technique for Generating and Observing Chemiluminescence in a Biological Setting

Published on: March 9, 2017

8.6K

Related Experiment Videos

Last Updated: Sep 29, 2025

Bioluminescence Imaging of NADPH Oxidase Activity in Different Animal Models
10:42

Bioluminescence Imaging of NADPH Oxidase Activity in Different Animal Models

Published on: October 22, 2012

16.4K
Fast and Specific Assessment of the Halogenating Peroxidase Activity in Leukocyte-enriched Blood Samples
05:17

Fast and Specific Assessment of the Halogenating Peroxidase Activity in Leukocyte-enriched Blood Samples

Published on: July 28, 2016

10.4K
A Novel Technique for Generating and Observing Chemiluminescence in a Biological Setting
08:57

A Novel Technique for Generating and Observing Chemiluminescence in a Biological Setting

Published on: March 9, 2017

8.6K

Area of Science:

  • Biochemistry
  • Enzymology
  • Chemiluminescence

Background:

  • Peroxidase enzymes catalyze reactions producing light through luminol dioxygenation.
  • Understanding reaction kinetics is crucial for quantifying enzyme activity.
  • Different peroxidases, like HRP, MPO, and EPO, exhibit distinct catalytic mechanisms.

Purpose of the Study:

  • To determine the reaction order of reactants in luminol luminescence for common and haloperoxidases.
  • To differentiate the kinetic behavior of horseradish peroxidase (HRP) from myeloperoxidase (MPO) and eosinophil peroxidase (EPO).
  • To elucidate the mechanistic differences between radical and non-radical peroxidase actions.

Main Methods:

  • Quantification of light emission from luminol dioxygenation.
  • Determination of reaction velocity dependence on reactant concentrations (H2O2, halide, luminol).
  • Comparative kinetic analysis of HRP, MPO, and EPO under varying pH and substrate conditions.

Main Results:

  • HRP-catalyzed luminol luminescence is first order for H2O2, while MPO and EPO are second order.
  • MPO and EPO show first-order kinetics for halide (Cl- or Br-) and luminol, unlike HRP.
  • Haloperoxidase action requires acidic conditions and involves non-radical oxidation, distinct from HRP's alkaline, radical mechanism.

Conclusions:

  • Luminol luminescence provides a sensitive method to distinguish between different peroxidase types based on their unique reaction kinetics.
  • The distinct kinetic profiles of HRP, MPO, and EPO highlight their different catalytic mechanisms and substrate specificities.
  • Haloperoxidase activity, particularly with chloride and bromide, can be specifically quantified in acidic environments using luminol luminescence.